Reference - Detail
| RRC ID | 38798 |
|---|---|
| Author | Tomita M, Ohkura N, Ito M, Kato T, Royce PM, Kitajima T. |
| Title | Biosynthesis of recombinant human pro-alpha 1(III) chains in a baculovirus expression system: production of disulphide-bonded and non-disulphide-bonded species containing full-length triple helices. |
| Journal | Biochem J |
| Abstract |
We have investigated the expression of human procollagen III by insect cells infected with a recombinant baculovirus carrying cDNA for the pro-alpha1(III) chain of type-III collagen. A high level of expression was obtained, and a small proportion of the heterologously expressed pro-alpha1(III) chains formed normally disulphide-bonded procollagen III, which was secreted into the culture medium. This species displayed a melting temperature (Tm) of approx. 38 degrees C as assessed by its resistance to digestion by a mixture of trypsin and chymotrypsin, slightly lower than that of 39.5 degrees C for procollagen III synthesized by cultured human dermal fibroblasts, and reflected a slight degree of under-hydroxylation of prolyl residues. This is possibly a consequence of the lower incubation temperature of insect cells, or of an insufficiency of prolyl hydroxylase activity within them. A significant proportion of the expressed chains formed trimeric molecules of similar thermal stability containing an apparently full-length triple-helical region, but were not disulphide-bonded and not secreted. In addition to providing a source of recombinant human procollagen III, the system promises to be useful in the study of procollagen chain association and subsequent folding. |
| Volume | 312 ( Pt 3)(Pt 3) |
| Pages | 847-53 |
| Published | 1995-12-15 |
| DOI | 10.1042/bj3120847 |
| PMID | 8554530 |
| PMC | PMC1136192 |
| MeSH | Amino Acids / analysis Animals Ascorbic Acid / pharmacology Baculoviridae / genetics* Base Sequence Cell Line Chymotrypsin / metabolism Disulfides / chemistry* Drug Stability Gene Expression* Hot Temperature Humans Molecular Sequence Data Procollagen / chemistry* Procollagen / genetics* Procollagen / metabolism Protein Conformation Recombinant Proteins / biosynthesis Spodoptera / metabolism Trypsin / metabolism |
| IF | 4.097 |
| Times Cited | 30 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 6 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | Sf9(RCB0563) |